Synthesis of macrocyclic cancer chemotherapy agents and methods of use

ABSTRACT

Herein are described a process for forming a quaternary carbon useful in the preparation of macrolactones, an enantioselective synthesis of (+)-peloruside A, and methods for treating a patient in need of relief from cancer or a cancer-related disease. The described processes are useful for preparing compounds containing quaternary carbons, including structural analogs and derivatives of peloruside A.

GOVERNMENT RIGHTS

This invention was made with Government support under Grant No. NIGMS53386, awarded by the National Cancer Institute and the NationalInstitutes of Health. The Government has certain rights in theinvention.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a U.S. national application under 35 U.S.C. §371(b)of International Application Serial No. PCT/US2009/030597 filed Jan. 9,2009, which claims priority under 35 USC §119(e) to U.S. ProvisionalApplication Ser. No. 61/020,498, filed Jan. 11, 2008, the disclosure ofwhich is incorporated herein by reference

BACKGROUND AND SUMMARY

Peloruside A (1), a 16-membered macrolide antitumor agent was firstisolated by West and Northcote from the New Zealand marine sponge,Mycale hentscheli (West, L. M.; Northcote, P. T.; Battershill, C. N. JOrg Chem 2000, 65, 445). It has shown potent antitumor activity againstP388 murine leukemia cells with an IC₅₀ value of 10 ng/mL. Peloruside Ais a microtubule stabilizing agent and arrests cells in the G2-M phase(Hood, K. A.; West, L. M.; Rouwe, B.; Northcote, P. T.; Berridge, M. V.;Wakefield, S. J.; Miller, J. H. Cancer Res 2002, 62, 3356). However,like laulimalide, peloruside A binds to the non-taxoid site of tubulinand has shown to a synergistic effect with taxol (Pryor, D. E.; O'Brate,A.; Bilcer, G.; Diaz, J. F.; Wang, Y.; Wang, Y.; Kabaki, M.; Jung, M.K.; Andreu, J. M.; Ghosh, A. K.; Giannakakou, P.; Hamel, E. Biochemistry2002, 41, 9109). The intriguing structure, very low natural abundanceand important biology of peloruside A attracted immense interest insynthesis ((a) Paterson, I.; Di Francesco, M. E.; Kuhn, T. Org. Lett.2003, 5, 599; (b) Ghosh, A. K.; Kim, J.-H. Tetrahedron Letters 2003, 44,3967; (c) Ghosh, A. K.; Kim, J.-H. Tetrahedron Letters 2003, 44, 7659;(d) Liu, B.; Zhou, W. S. Org. Lett. 2004, 6, 71). Thus far, De Brabanderet al. and subsequently Taylor and co-workers have achieved the totalsynthesis of peloruside A ((a) Liao, X.; Wu, Y.; De Brabander, J. K.Angew Chem Int Ed Engl 2003, 42, 1648; (b) Jin, M.; Taylor, R. E. Org.Lett. 2005, 7, 1303). Even so, additional synthetic procedures areuseful to ensure the supply of peloruside A, and also to provide a routeto analogs and derivatives of this important molecule.

Described herein is a process for forming a quaternary carbon, theprocess comprising the steps of; reacting a compound containing a2-alkyl-2-ene-1-one moiety with a source of nucleophilic hydride; andadding a compound containing an aldehyde to form the quaternary carbon.The process may be used to prepare a wide variety of chemical compounds.In one illustrative embodiment, the process may be used for thepreparation of a macrolactone such as, but not limited to, a peloruside,a mycalamide, a bryostatin, an epothilone, and the like.

Also described herein is a process for preparing a peloruside, includingpeloruside A and analogs and derivatives thereof. The process comprisesthe step of preparing an aldol intermediate that is subsequentlyconverted into the peloruside. The compounds described herein are usefulfor treating a patient in need of relief from cancer, a cancer-relateddisease, or a disease linked to the presence of a population ofpathogenic cells.

DETAILED DESCRIPTION

As shown below, described herein is a synthetic process that includesthe assembly of fragments I and II by a stereoselective aldol reaction,followed by a macrolactonization of the corresponding carboxylic acid atC-1 and hydroxyl group at C-16. Without being bound by theory, it isbelieved that the presence of the gem-dimethyl group at C-10, makes theC-7 to C-11 lactol segment of peloruside A sterically very hindered. Ithas been reported that aldol reactions involving gem-dimethyl ketone andaldehyde often result in poor yield and side reactions, as described byLiu, B.; Zhou, W. S. Org. Lett. 2004, 6, 71. The foregoing citation,along with all other citations disclosed herein, are incorporated hereinby reference. Both published peloruside A syntheses utilized methylketone aldol reactions to avoid this reported problem ((a) Liao, et al,2003; Jin, et al, 2005). In contrast, it has been discovered herein thatthe process described herein provides for the installation of the C-10gem-dimethyl and C-11 hydroxyl group by an efficient reductiveenolization of enone II followed by reaction with aldehyde I.

Retrosynthetic Route of the Processes Described Herein

In one embodiment, a process for forming a quaternary carbon isdescribed, the process comprises the steps of (a) reacting a compoundcontaining a 2-alkyl-2-ene-1-one moiety with a source of nucleophilichydride; and (b) adding a second compound containing an aldehyde to formthe quaternary carbon.

In another embodiment, the process of forming a compound wherein thecompound contains a quaternary carbon is described, where the processcomprises the steps of (a) reacting a compound containing a2-alkyl-2-ene-1-one moiety with a source of nucleophilic hydride; and(b) adding a second compound containing an aldehyde to form thequaternary carbon.

In another embodiment, the processes described above wherein thecompound containing a quaternary carbon is a macrolactone are described.

In another embodiment, any of the processes described above wherein themacrolactone is a bryostatin, a peloruside, a mycalamide or anepothilone are described.

In another embodiment, any of the processes described above wherein themacrolactone modulates microtubule assembly are described.

In another embodiment, any of the processes described above wherein themacrolactone has the formula

wherein:

R¹ is

where * shows the point of attachment and R8 and R12 are independentlyhydrogen, alkyl, heteroalkyl, arylalkyl, heteroarylalkyl, or an oxygenprotecting group;

R², R³, R⁴, R⁵, R⁶ and R⁷ are each independently selected from the groupconsisting of hydrogen, alkyl, hydroxy, and alkoxy; and X¹ is hydrogen,hydroxy, alkoxy or together with the carbon to which it is attachedforms a carbonyl or an oxime are described.

In another embodiment, any of the processes described above wherein themacrolactone has the formula

wherein:

R¹ is

where * shows the point of attachment;

R², R³, R⁴, R⁵, R⁶ and R⁷ are each independently selected from the groupconsisting of hydrogen, alkyl, hydroxy, and alkoxy; X¹ is hydrogen,hydroxy, alkyl, alkoxy or together with the carbon to which it isattached forms a carbonyl or an oxime; and

R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl, arylalkyl,heteroarylalkyl, or an oxygen protecting group are described.

In another embodiment, any of the processes described above wherein themacrolactone has the formula

wherein

R¹ is

where * shows the point of attachment;

R², R³, R⁴, R⁵, R⁶ and R⁷ are each independently selected from the groupconsisting of hydrogen, alkyl, hydroxy, and alkoxy; X¹ is hydrogen,hydroxy, alkyl, alkoxy or together with the carbon to which it isattached forms a carbonyl or an oxime; and

R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl, arylalkyl,heteroarylalkyl, or an oxygen protecting group are described.

In another embodiment, any of the processes described above wherein R³is methoxy and R⁷ is methoxy are described.

In another embodiment, any of the processes described above wherein R⁶is hydrogen is described. In another embodiment, any of the processesdescribed above wherein R² is hydrogen are described.

In another embodiment, any of the processes described above wherein R²is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydroxy;and

R¹ is

where * shows the point of attachment and R⁸ and R¹² are independentlyhydrogen, alkyl, heteroalkyl, arylalkyl, heteroarylalkyl, or an oxygenprotecting group are described.

In another embodiment, any of the processes described above wherein R²is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydrogen, hydroxy,alkyl, such as methyl or ethyl, and the like, or alkoxy, such as methoxyor ethoxy, and the like; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, any of the processes described above wherein R²is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydrogen,or epi-hydroxy, alkoxy, such as methoxy or ethoxy, and the like, oralkyl, such as methyl or ethyl, and the like or X¹ and the attachedcarbon form a carbonyl; and R¹ is 4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-ylare described.

In another embodiment, any of the processes described above wherein R²is hydrogen, hydroxy, or alkyl, such as methyl or ethyl, and the like;R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, any of the processes described above wherein R²is hydroxy; R⁴ is hydrogen, hydroxy, alkoxy, such as methoxy or ethoxy,and the like, or alkyl, such as methyl or ethyl, and the like; R⁵ ishydroxy; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, any of the processes described above wherein R²is hydroxy; R⁴ is methoxy; R⁵ is hydrogen, hydroxy, alkoxy, such asmethoxy or ethoxy, and the like, or alkyl, such as methyl or ethyl, andthe like; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, any of the processes described above wherein thesource of nucleophilic hydride is lithium (sec-butyl)₃borohydride aredescribed.

In one embodiment analogs of peloruside may be prepared by the processesdescribed herein.

In another aspect, the C7 and C8 hydroxyl groups in 2 may provided byasymmetric dihydroxylation while the C5 center may be setup by Brownallylation (Jadhav, P. K.; Bhat, K. S.; Perumal, P. T.; Brown, H. C. J.Org. Chem. 1986, 51, 432) and the C2 and C3 chiral centers may bederived from known compound 4. Aldehyde 3 may be obtained fromasymmetric allylation of the corresponding aldehyde derived from knowncompound 5 (Ghosh, A. K.; Kim, J.-H. Tetrahedron Letters 2003, 44,7659).

Conditions: (a) NaH, PMBCl, 23° C. 78%; (b) Ph₃P, I₂, Imidazole, 0° C.,88%; (c) CH₂═CHMgBr, CuI, HMPA, −30° C. to 0° C., 85%; (d) 10% HCl,MeOH, 23° C.; (e) CH₃CN, NaHCO₃, I₂, 0° C. to 23° C., 82% (2 steps); (f)Me₃O⁺BF₄ ⁻, proton sponge, 23° C., 87%; (g) 95% EtOH, Zn; (h) MOMC1,DIPEA, 23° C., 88% (2 steps); (i) NMO, OsO₄, Acetone/H₂O, 0° C.; (j)NaIO₄, THF/H₂O, 23° C.; (k) (+)-Ipc₂BOMe, CH₂═CHCH₂MgBr, 23° C.,aldehyde, −78° C., 2 h, NaOH, H₂O₂, 77% (3 steps); (1) TBSCl, imidazole,DMAP, 96%; (m) (o-cresol)₂P(O)CH₂CO₂Et, NaH, NaI, 0° C., aldehyde, −78°C. to −50° C., 89% (3 steps); (n) AD-mix-α, CH₃SO₂NH₂, ^(t)BuOH—H₂O, 0°C., 72 h, 97%; (O)CH₂═C(OMe)Me, PPTS, 94%; (p) Dibal-, −78° C., 96%; (q)CH₂═C(Me)MgBr, THF, 0° C., 86%; (r) Dess-Martin, 23° C., 90%.

One illustrative embodiment of the processes described herein is shownin Scheme 1. The synthesis of C1-C11 segment 2 commences with thecommercially available (−)-2,3-O-isopropylidene-D-threitol 4. It isconverted to iso-propylidene derivative 6 in a three step sequenceinvolving; (1) mono benzylation of 4 with sodium hydride and PMBCl; (2)conversion of alcohol to iodide and (3) allylation of the resultingiodide with Grignard reagent. Acid-catalyzed removal of isopropylidenegroup followed by iodoetherfication with iodine in the presence ofsodium bicarbonate and methylation of the C3 hydroxyl group withtrimethyloxonium tetrafluoroborate (Earle, M.; Fairhurst, R.; Giles, R.;Heaney, H. Synlett 1991, 728) affords iodide 7. Reductive cleavage ofthe iodoether followed by protection of alcohol as its MOM etherprovides 8. Conversion of the terminal olefin to aldehyde (Yu, W.; Mei,Y.; Kang, Y.; Hua, Z.; Jin, Z. Org. Lett. 2004, 6, 3217) and asymmetricallylation followed by TBS protection and furnishes 10. The terminalolefin is converted to aldehyde as described above and Horner-Emmonsolefination of the resulting aldehyde furnishes the Z olefin 11selectively (Z:E 7:1, 89% in 3 steps). Sharpless asymmetricdihydroxylation (Kolb, H. C.; VanNieuwenhze, M. S.; Sharpless, K. B.Chem. Rev. 1994, 94, 2483) of the pure Z-olefin proceeds in excellentyield (97%) and diastereoselectivity (dr 6.3:1 by 1H NMR analysis).Protection of the diol as the isopropylidene derivative yields the ester12. Dibal-H reduction of the ester followed by addition of the Grignardreagent and subsequent Des s-Martin oxidation of the resulting alcoholaccomplishes the synthesis of the enone segment 2.

The synthesis of C11-C24 segment 3 is shown in Scheme 2. Homoallylicalcohol 5 is synthesized utilizing chiral imide 13 as describedpreviously (Ghosh, A. K.; Kim, J.-H. Tetrahedron Letters 2003, 44,7659). The hydroxyl group is protected as its TES ether. Oxidativecleavage of the terminal olefin provides aldehyde which is exposed toasymmetric allylation (Jadhav, et al; 1986) to furnish alcohol 15diastereoselectively (dr 5:1 by 1H NMR analysis). Alcohol 15 isconverted to methyl ether as described above and oxidative cleavage ofthe resulting olefin provides aldehyde 3 in good yield. With thesyntheses of enone 2 and aldehyde 3, our synthetic strategy calls forthe assembly of these segments by reductive enolization of enone 2followed by aldol addition to aldehyde 3. Thus, reaction of 2 with 1.1equivalent of L-selectride at −78° C. for 10 min provides thecorresponding enolate. Reaction of this enolate with aldehyde 3 at −78°C. for 1 h affords the aldol product 17 and its diastereomer, 17a, as a4:1 mixture in 92% isolated yield. The diastereomers are readilyseparated by silica gel chromatography. This aldol protocol is practicaland the efficiency of this process is significantly improved compared todirect aldol reaction with a related ketone enolate and aldehyde (LDAinduced a-dimethyl ketone aldol coupling results in a 29% yield and 40%loss of starting material).

Conditions: (a) TESOTf, 2,6-lutidine, 90%; (b) NMO, OsO₄, Acetone/H₂O,0° C.; (c) Pb(OAc)₄, CH₂Cl₂; (d) (−)-Ipc₂BOMe, Allyl magnesium bromide,23° C., then −78° C., 2 h, NaOH, H₂O₂, 61% (3 steps); (e) Me₃OBF₄,proton sponge, 23° C., 98%; (f) NMO, OsO₄, Acetone/H₂O, 0° C.; (g)Pb(OAc)₄, CH₂Cl₂, 74% (2 steps); (h) L-selectride, Et₂O, −78° C., 10min, then 3, −78° C., 1 h; 92%.

The subsequent elaboration to macrolactone and synthesis of peloruside Ais shown in Scheme 3. The TES group of 17 is selectively removed byreaction with a catalytic amount of DDQ in aqueous THF. Subsequentexposure of resulting PMB ether to an excess of DDQ in the presence ofpH 7 buffer removes the PMB group. TPAP oxidation of 18 selectivelyoxidizes the primary alcohol to an aldehyde, which is oxidized withsodium chlorite to the carboxylic acid. The resulting acid is subjectedto Yamaguchi lactonization (Inanaga, J. H., K.; Saeki, H.; Katsuki, T.and Yamaguchi, M., Bull. Chem. Soc. Jpn. 1979, 52, 1989) protocol with2,4,6-trichlorobenzoyl chloride in the presence of DMAP to provide thecorresponding macrolactone 19 in good yield. Macrolactone 19 isconverted to synthetic (+)-peloruside A as follows: deprotection of theTBS and isopropylidene groups with 1M aqueous HCl to provide ahemiketal, selective methylation of the equatorial hydroxyl group withtrimethyloxonium tetrafluoroborate, removal of the benzyl group bytransfer hydrogenation conditions and removal of MOM group by exposureto aqueous 4N HCl (Removal of isopropylidene group results in anequilibrium mixture of macrocycle and hemi-ketal. Final chromatographicpurification is made after the final step).

Conditions: (a) DDQ, THF/H₂O, 23° C., 2 h, then DDQ, CH₂Cl₂, pH 7, 5 h,70%; (b) TPAP, NMO, CH₂Cl₂, 4 Å MS, 0° C.; (c) NaClO₂, NaH₂PO₄, H₂O,tBuOH, 2-methyl-2-butene; 23° C., 52% (2 steps) (d) 2,4,6-Cl₃-C₆H₂COCl,Et₃N, PhMe, then DMAP, PhMe, 23° C., 24 h, 64%; (e) 1M HCl, THF, 23° C.,8 h; (f) Me₃OBF₄, 2,6-di-tert-butylpyridine, 0° C.; 73% (2 steps) (g)Pd/C, HCOOH, MeOH, 23° C., 1 h; (h) 4N aq. HCl, THF, 23° C., 3.5 h; 50%(2 steps).

In the example where the compound was (+)-peloruside A, spectral data(1H and 13C NMR) of synthetic compound (1) was identical to thatreported for the naturally occurring compound.

In another embodiment an intermediate compound of structure

wherein R¹ is

R² is hydrogen, methyl, alkyl, hydroxy or alkoxy; R³ is hydrogen,methyl, ethyl, alkyl, methoxy, or alkoxy; R⁴, R⁵ and R⁷ are eachindependently selected from the group consisting of hydrogen, methyl,ethyl, alkyl, hydroxy, methoxy and alkoxy; R⁸ and R¹² are in eachinstance independently selected from the group consisting of hydrogen,alkyl, alkylaryl and oxygen protecting group; R⁹ and R¹⁰ are each anindependently selected oxygen protecting group; which is useful for themanufacture of a compound of the structure

wherein:

R¹, R², R³, R⁴, R⁵, and R⁷, R⁸, R⁹ and R¹⁰ are as defined above; R⁶ ishydrogen, methyl, alkyl, hydroxy or alkoxy; X¹ is hydroxy, alkyl,alkoxy, or together with the carbon to which it is attached to form acarbonyl or and oxime; is synthesized by reacting a compound ofstructure

wherein R², R³, R⁴, R⁵, R⁹ and R¹⁰ are as defined above; with a sourceof nucleophilic hydride and adding to the mixture a compound ofstructure

wherein; R¹, R⁷ and R¹¹ are defined above to yield the intermediatecompound.

In one embodiment compounds of the formula

or a pharmaceutically acceptable salt thereofwherein:

R¹ is

where * shows the point of attachment;

R², R³, R⁴, R⁵, R⁶ and R⁷ are each independently selected from the groupconsisting of hydrogen, alkyl, hydroxy, and alkoxy; X¹ is hydrogen,hydroxy, alkoxy, alkyl or together with the carbon to which it isattached forms a carbonyl or an oxime; and

R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl, arylalkyl,heteroarylalkyl, or an oxygen protecting group;

providing that the compound is not (+)-peloruside A or (−)-peloruside A;and providing that a) when R²=R⁶=X¹=hydroxy and R³=R⁴=OMe, then R⁵ ishydroxy; b) when R⁵=R⁶=X¹=hydroxy and R³=R⁴=OMe, then R² is notmethoxymethoxy; and c) R² and R³ are not both hydrogen are described.

In another embodiment, the compounds described in the precedingembodiment having the formula

wherein

R¹ is

where * shows the point of attachment;

R², R³, R⁴, R⁵, R⁶ and R⁷ are each independently selected from the groupconsisting of hydrogen, alkyl, hydroxy, and alkoxy; X¹ is hydrogen,hydroxy, alkyl, alkoxy or together with the carbon to which it isattached forms a carbonyl or an oxime; and

R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl, arylalkyl,heteroarylalkyl, or an oxygen protecting group are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R³ is methoxy and R⁷ is methoxy are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R⁶ is hydrogen are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydrogen are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R³ is methoxy, R⁷ is methoxy, and R⁶ is hydrogen are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R³ is methoxy, R⁷ is methoxy, and R² is hydrogen are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R³ is methoxy, R⁷ is methoxy, R⁶ is hydrogen, and R² is hydrogenare described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹is hydroxy; and

R¹ is

where * shows the point of attachment and R⁸ and R¹² are independentlyhydrogen, alkyl, heteroalkyl, arylalkyl, heteroarylalkyl, or an oxygenprotecting group are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydrogen,hydroxy, alkyl, such as methyl or ethyl, and the like, or alkoxy, suchas methoxy or ethoxy, and the like; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹is hydrogen, or epi-hydroxy, or X¹ and the attached carbon form acarbonyl; and R¹ is 4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl aredescribed.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydrogen, hydroxy, or alkyl, such as methyl or ethyl, andthe like; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydroxy;and R¹ is 4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydroxy; R⁴ is hydrogen, hydroxy, alkoxy, such as methoxyor ethoxy, and the like, or alkyl, such as methyl or ethyl, and thelike; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In another embodiment, the compounds of any of the preceding embodimentswherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydrogen, hydroxy, alkoxy,such as methoxy or ethoxy, and the like, or alkyl, such as methyl orethyl, and the like; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl are described.

In some variations of the embodiments described above, the term alkylrefers to C₁-C₄ alkyl. In some variations of the embodiments describedabove, the term alkoxy refers to C₁-C₄ alkoxy.

In another illustrative embodiment, a method for treating a patientsuffering from or in need of relief from cancer, a cancer-relateddisease or a disease linked to the presence of a population ofpathogenic cells is described, the method comprising administering tothe patient a therapeutically effective amount of a compositioncomprising any of the compounds described herein.

In another embodiment, the compounds and methods described herein areused in conjunction with taxol, paclitaxol, and the like, or anothercompound that binds to taxol site.

The following non-limiting examples are described, each of which may beprepared by the processes described herein:

where R¹ is

where R⁴=OH, CH₃, H, OCH₃ or CH₂CH₃; and R⁵=OH, CH₃, H, OCH₃ or CH₂CH₃;

where R⁶=OH, H, CH₃;

where X¹=OH, ═O;

where R⁷=OH, OCH₃, CH₃, CH₂CH₃, H;

where R²=H, OH, CH₃; and

where R³=H, OCH₃, CH₂CH₃, CH₃.

In another embodiment, compounds of the present invention can beprepared and administered in a wide variety of oral, parenteral andtopical dosage forms. Thus, the compounds of the present invention canbe administered by injection (e.g. intravenously, intramuscularly,intracutaneously, subcutaneously, intraduodenally, orintraperitoneally). Also, the compounds described herein can beadministered by inhalation, for example, intranasally. Additionally, thecompounds of the present invention can be administered transdermally. Itis also envisioned that multiple routes of administration (e.g.,intramuscular, oral, transdermal) can be used to administer thecompounds of the invention. Accordingly, the present invention alsoprovides pharmaceutical compositions comprising one or morepharmaceutically acceptable diluents, carriers or excipients and one ormore compounds of the invention.

For the treatment of cancer and disease linked to the presence of apopulation of pathogenic cells, illustratively the compounds describedherein may be formulated in a therapeutically effective amount inconventional dosage forms, including one or more carriers, diluents,and/or excipients. Such formulation compositions may be administered bya wide variety of conventional routes in a wide variety of dosageformats, utilizing art-recognized products. See generally, Remington'sPharmaceutical Sciences, (16th ed. 1980). It is to be understood thatthe compositions described herein may be prepared from isolatedcompounds described herein or from salts, solutions, hydrates, solvates,and other forms of the compounds described herein. It is also to beunderstood that the compositions may be prepared from various amorphous,non-amorphous, partially crystalline, crystalline, and/or othermorphological forms of the compounds described herein.

In making the formulations of the compounds described herein, atherapeutically effective amount of the compounds herein described, inany of the various forms described herein, may be mixed with anexcipient, diluted by an excipient, or enclosed within such a carrierwhich can be in the form of a capsule, sachet, paper, or othercontainer. Excipients may serve as a diluent, and can be solid,semi-solid, or liquid materials, which act as a vehicle, carrier ormedium for the active ingredient. Thus, the formulation compositions canbe in the form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solidor in a liquid medium), ointments, soft and hard gelatin capsules,suppositories, sterile injectable solutions, and sterile packagedpowders. The compositions may contain anywhere from about 0.1% to about99.9% active ingredients, depending upon the selected dose and dosageform. Some examples of suitable excipients include lactose, dextrose,sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate,alginates, tragacanth, gelatin, calcium silicate, microcrystallinecellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxybenzoates; sweetening agents; and flavoring agents. Thecompositions can be formulated so as to provide quick, sustained ordelayed release of the active ingredient after administration to thepatient by employing procedures known in the art. It is appreciated thatthe carriers, diluents, and excipients used to prepare the compositionsdescribed herein are advantageously GRAS (Generally Regarded as Safe)compounds.

The unitary daily dosage of the compounds described in the invention canvary significantly depending on the host condition, the disease statebeing treated, the molecular weight of the conjugate, its route ofadministration and tissue distribution, and the possibility of co-usageof other therapeutic treatments such as radiation therapy. The effectiveamount to be administered to a patient is based on body surface area,patient weight, and physician assessment of patient condition. Aneffective dose can range from about 1 ng/kg to about 50 mg/kg, fromabout 0.10 μg/kg to about 10 mg/kg, from about 1 μg/kg to about 5 mg/kg,and from about 10 μg to about 1 mg/kg.

Any effective regimen for administering the composition comprising acompound of the invention can be used. For example, the compositioncomprising a compound of the invention can be administered as singledoses, or it can be divided and administered as a multiple-dose dailyregimen. Further, a staggered regimen, for example, one to three daysper week can be used as an alternative to daily treatment, and for thepurpose of defining this invention such intermittent or staggered dailyregimen is considered to be equivalent to every day treatment and withinthe scope of this invention.

The compounds described herein may contain one or more chiral centers,or may otherwise be capable of existing as multiple stereoisomers.Accordingly, it is to be understood that the present invention includespure stereoisomers as well as mixtures of stereoisomers, such asenantiomers, diastereomers, and enantiomerically or diastereomericallyenriched mixtures. The compounds described herein may be capable ofexisting as geometric isomers. Accordingly, it is to be understood thatthe present invention includes pure geometric isomers or mixtures ofgeometric isomers.

It is appreciated that the compounds described herein may exist inunsolvated forms as well as solvated forms, including hydrated forms. Ingeneral, the solvated forms are equivalent to unsolvated forms and areencompassed within the scope of the present invention. The compounds ofthe present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

It is also appreciated that in the foregoing embodiments, certainaspects of the compounds are presented in the alternative, such asselections for any one or more of X¹, R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R⁸,R⁹, R¹⁰, R¹¹, R¹². It is therefore to be understood that variousalternate embodiments of the invention include individual members ofthose lists, as well as the various subsets of those lists. Each ofthose combinations are to be understood to be described herein by way ofthe lists.

Additional features of the present invention will become apparent tothose skilled in the art upon consideration of the following descriptionof illustrative embodiments for carrying out the invention.

Methods

General Experimental Methods. All moisture sensitive reactions werecarried out under nitrogen or argon atmosphere. Anhydrous solvents wereobtained as follows: THF, diethyl ether and benzene, distilled fromsodium and benzophenone; dichloromethane, pyridine, triethylamine, anddiisopropylethylamine, distilled from CaH₂. All other solvents were HPLCgrade. Column chromatography was performed with 240-400 mesh silica gelunder low pressure of 5-10 psi. TLC was carried out with silica gelplates. ¹H and ¹³C NMR spectra were recorded on 500 or 400 MHzspectrometers. Infrared spectra were recorded on a FTIR instrument.

Example 1

Olefin (6). To the solution of (−)-2,3-O-isopropylidene-D-threitol (4)(5.5 g, 34 mmol) in THF (60 mL) was added NaH (60%, 1.49 g, 37 mmol) at0° C., and the reaction was warmed up to 23° C. over 1 h. PMBCl (4.85mL, 34 mmol) was added at 23° C. and the reaction mixture was stirredfor 1.5 h and was quenched with aqueous NH₄Cl. The mixture was extractedwith ether and the organic layer was washed with water and brine. Theresulting mixture was dried over Na₂SO₄ and concentrated under reducedpressure. Purification by column chromatography provided product (7.5 g,78%). To a solution of the mono-PMB protected product (87.3 g, 0.31 mol)in THF (700 mL) was added imidazole (52.6 g, 0.77 mol), Ph₃P (122 g,0.46 mol) and iodine (118 g, 0.46 mol) at 0° C. successively. Theresulting mixture was warmed up to 23° C. over 2 h and stirred overnightand then quenched by 10% aqueous Na₂S₂O₃. The mixture was extracted withether and the organic layer was washed with water and brine. Theresulting mixture was dried over Na₂SO₄ and concentrated under reducedpressure. Purification by column chromatography provided (106 g, 88%)iodide as a colorless oil. [α]²³ _(D)=+12.2 (c 2.21, CHCl₃); IR (thinfilm, cm⁻¹) 2986, 1612, 1514, 1091, 821; ¹H NMR (500 MHz, CDCl₃), δ 7.23(2H, d, J=6.5 Hz), 6.88 (2H, d, J=6.5 Hz), 4.51 (2H, s), 3.94 (1H, dt,J=2.5, 5.0 Hz), 3.83 (1H, dt, J=3.0, 7.5 Hz), 3.81 (3H, s), 3.63 (1H,dd, 10.0, 5.0 Hz), 3.59 (1H, dd, J=10.0, 5.0 Hz), 3.33 (3H, dd, J=5.0,10.5 Hz), 3.26 (3H, dd, J=5.5, 10.5 Hz), 1.46 (3H, s), 1.41 (3H, s); ¹³CNMR (125 MHz, CDCl₃) δ 159.3, 129.9, 129.4, 113.9, 109.8, 80.1, 77.7,73.3, 70.2, 55.3, 27.4, 27.3, 6.5; MS (EI, m/z) [M]⁺392.04.

To a solution of the above iodide (34.3 g, 87 mmol) in THF (100 mL) wasadded HMPA (62 mL) and CuI (3.4 g, 17.2 mmol) at 23° C. The resultingmixture was cooled to −30° C. and vinylmagnesium bromide (173 mL, 1M inTHF, 173 mmol) was added dropwise at that temperature over 1 h. Theresulting mixture was stirred at −30° C. for 1 h and then warmed up to10° C. and then quenched with aqueous NH₄Cl. The organic layer wasseparated and the aqueous layer was extracted with Et₂O, the combinedorganic layer was washed with brine, dried over Na₂SO₄ and concentratedin vacuo. Column chromatography provided product 6 (21.6 g, 85%). [α]²³_(D)=+ 15.0 (c 3.05, CH₂Cl₂); IR (thin film, cm⁻¹) 3075, 2985, 2933,2906, 2864, 2838, 1613, 1514, 1369, 1248, 1172, 1086, 1036, 917; ¹H NMR(400 MHz, CDCl₃) δ 7.25 (2H, d, J=8.5 Hz), 6.88 (2H, d, J=8.7 Hz), 5.82(1H, m), 5.06-5.13 (2H, m), 4.53 (1H, d, J=11.8 Hz), 4.49 (1H, d, J=11.8Hz), 3.86 (2H, m), 3.79 (3H, s), 2.36 (2H, m), 1.41 (3H, s), 1.40 (3H,s); ¹³C NMR (100 MHz, CDCl₃) δ 159.1, 133.7, 130.0, 129.2, 117.5, 113.7,108.8, 79.5, 77.3, 73.1, 70.0, 55.2, 37.3, 27.1, 26.9; MS (ESI, m/z)[M+Na]⁺315.0; HRMS (ESI) [M+Na]⁺ calcd for C₁₇H₂₄O₄Na 315.1572, found315.1571.

Example 2

Methyl ether (7) To the obtained olefin 6 (31.5 g, 108 mmol) in methanol(500 mL) was added 10% HCl (66 mL) at 0° C. and the mixture was stirredat 23° C. for 12 h. The reaction was then quenched with Na₂CO₃ (11.4 g,108 mmol) and concentrated to give the crude diol which was used for thenext step without further purification. The crude diol (57 g, 226 mmol)was dissolved in MeCN (1.3 L), and NaHCO₃ (109 g, 1.3 mol) and iodine(125 g, 490 mmol) was added successively at 0° C. The resulting mixturewas warmed up to 23° C. over 3 h and then quenched by 10% aqueousNa₂S₂O₃ and extracted with ethyl acetate. The combined organic layer waswashed with brine and concentrated in vacuo. Column chromatographyprovided product (70.4 g, 82.4% for 2 steps) as a solid. mp 60-62° C.;[α]²³ _(D)−24 (c 2.64, CHCl₃); ¹H NMR (500 MHz, CDCl₃) δ 7.30 (2H, m),6.87 (2H, m), 4.53-4.48 (3H, m), 4.26 (0.75H, m), 4.15 (0.75H, dd,J=4.0, 9.0 Hz), 4.09 (0.25H, m), 3.95 (0.25H, dd, J=4.5, 9.0 Hz), 3.75(2H, m), 3.36 (0.5H, m), 3.33-3.27 (1.5H, m), 3.07 (1H, s), 2.35 (0.25H,ddd, J=14.0, 7.0, 7.0 Hz), 2.20 (0.75H, dd, J=5.5, 14.0 Hz), 1.86 (1H,m); ¹³C NMR (125 MHz, CDCl₃) 159.5, 129.6, 129.5, 114.0, 109.8, 81.7,81.0, 78.0, 74.0, 73.6, 73.0, 68.7, 68.6, 55.3, 42.3, 40.8, 11.2, 10.7;IR (thin film, cm⁻¹) 3440, 2931, 1612, 1513, 1071, 820; To the solutionof the alcohol (24.5 g, 64.8 mmol) in CH₂Cl₂ (240 mL) was added protonsponge (17.3 g, 81 mmol) and Me₃OBF₄ (11.7 g, 79 mmol) at 0° C. Thereaction mixture was then warmed up to 23° C. and stirred over night.The solid was removed by filtration and the organic layer was washedwith aqueous NaHCO₃, water and brine, and dried over Na₂SO₄ andconcentrated in vacuo. Column chromatography provided the methyl ether 7(22 g, 87%). It's a mixture (dr 4:1). [α]²³ _(D)-41.8 (c 1.83, CH₂Cl₂);IR (thin film, cm⁻¹) 2929, 2902, 2864, 2835, 1612, 1513, 1462, 1247,1085, 1034, 820; ¹H NMR (400 MHz, CDCl₃)

7.27 (2H, d, J=8.1 Hz), 6.87 (2H, d, J=8.6 Hz), 4.56 (1H, d, J=11.7 Hz),4.45 (0.2H, d, J=11.7 Hz), 4.44 (0.8H, d, J=11.7 Hz), 4.23 (0.8H, m),4.18 (1H, m), 4.06 (0.2H, m), 3.94 (0.8H, m), 3.89 (0.2H, m), 3.79 (3H,s), 3.71-3.67 (0.2H, m), 3.63 (0.8H, dd, J=5.0, 9.9 Hz), 3.58 (1H, dd,J=6.8, 10.1 Hz), 3.39-3.32 (1.5H, m), 3.30 (2.4H, s), 3.28 (0.6H, s),3.24-3.21 (1H, m), 2.35 (0.8H, m), 2.19 (0.2H, m), 2.04 (0.2H, ddd,J=2.3, 4.7, 13.7 Hz), 1.69 (0.8H, ddd, J=4.6, 9.1, 13.5 Hz); ¹³C NMR(100 MHz, CDCl₃)

159.1, 130.3, 129.3, 113.6, 82.6, 81.8, 81.7, 80.9, 78.3, 77.2, 76.8,72.9, 68.7, 68.3, 57.2, 57.1, 55.2, 37.8, 36.2, 11.0, 9.9; MS (ESI, m/z)[M=Na]⁺415.1; HRMS (ESI) m/z [M+Na]⁺ calcd for C₁₅H₂₁O₄INa 415.0382,found 415.0392.

Example 3

MOM ether (8) To a solution of the obtained methyl ether 7 (46.2 g, 118mmol) in 95% ethanol (610 mL) was added zinc dust (59.4 g, 910 mmol).The reaction mixture was stirred at 80° C. for 6 h and cooled down to23° C. The solid was removed by filtration and rinsed with EtOAc. Thefiltrate was concentrated in vacuo and purified by column chromatographyto provide the product (31 g, 99%) as a colorless oil. [α]²³ _(D)-8.04(c 2.55, CHCl₃); IR (thin film, cm⁻¹) 3457, 2930, 1716, 1612, 1514,1088, 824; ¹H NMR (500 MHz, CDCl₃) δ 7.25 (2H, d, J=8.4 Hz), 6.88 (2H,d, J=8.4 Hz), 5.82 (1H, m), 5.07 (2H, m), 4.48 (3H, s), 3.76 (3H, m),3.54-3.46 (2H, m), 3.39 (3H, s), 3.31 (1H, m), 2.38 (1H, m), 2.28 (1H,m); ¹³C NMR (125 MHz, CDCl₃) δ 159.3, 134.3, 130.1, 129.5, 117.5, 113.7,80.5, 73.1, 71.4, 70.7, 58.2, 55.3, 34.3; MS (ESI, m/z) [M+Na]⁺289.1. Toa stirred solution of the alcohol (31 g, 116 mmol) in CH₂Cl₂ (300 mL)was added diisopropylethylamine (69.7 mL 371 mmol) and MOMCl (23.6 mL,309 mmol) successively and the resulting mixture was stirred over nightand quenched with water. The organic layer was separated, washed withbrine, dried over Na₂SO₄ and concentrated in vacuo. Columnchromatography provided the MOM ether 8 (32 g, 88%) as a colorless oil.[α]²³ _(D) −13.6 (c 2.95, CHCl₃); ¹H NMR (500 MHz, CDCl₃) δ 7.25 (2H, d,J=9.0 Hz), 6.87 (2H, d, J=9.0 Hz), 5.83 (1H, m), 5.06 (2H, m), 4.78 (1H,d, J=6.5 Hz), 4.69 (1H, d, J=6.5 Hz), 4.78 (1H, A of AB, J=11.5 Hz),4.45 (1H, B of AB, J=11.7 Hz), 3.80 (s, 3H), 3.77 (1H, m), 3.63 (1H, dd,J=4.5, 9.5 Hz), 3.56 (1H, dd, J=6.0, 10.0 Hz), 3.42 (1H, m), 3.40 (3H,s), 3.38 (3H, s), 2.35 (1H, m), 2.28 (1H, m); ¹³C NMR (125 MHz, CDCl₃) δ159.2, 135.3, 130.3, 129.3, 117.1, 113.8, 97.1, 80.5, 73.0, 65.5, 58.7,55.7, 53.3, 34.5; MS (ESI, m/z) [M+Na]⁺333.0.

Example 4

Alcohol (9) To the solution of thus obtained MOM ether 8 (7 g, 22.5mmol) in acetone/H₂O (106 mL/13 mL) was added NMO (5.3 g, 45 mmol) andOsO₄ (2.5 w % in t-BuOH, 8.3 mL, 0.69 mmol). The resulting mixture wasstirred at 23° C. for 3 h and quenched with saturated aqueous NaHSO₃ (42mL). The solid was removed by filtration and the filtrate was extractedwith EtOAc. The combined organic layer was washed with brine, dried overNa₂SO₄ and concentrated in vacuo. To a solution of ⅓ of the crude diol(2.6 g, 7.49 mmol) in THF/H₂O (58 mL/14 mL) was added NaIO₄ (3.85 g, 18mmol) at 23° C. and stirred for 2 h. The solid was removed by filtrationand the filtrate was extracted with Et₂O. The combined organic layer waswashed with buffer solution (pH=7), water and brine, and dried overNa₂SO₄ and concentrated in vacuo. The aldehyde was used without furtherpurification. To the solution of (+)-Ipc₂BOMe (2.84 g, 9.0 mmol) in Et₂O(32 mL) was added allylmagnesium bromide_(S-5) (1M in Et₂O, 8.2 mL)dropwise at 0° C. The resulting mixture was warmed up to 23° C. over 1h. The solid was removed by filtration. And to the filtrate was addedthus obtained crude aldehyde in Et₂O (15 mL) via cannula at −78° C. over5 min, and the mixture was stirred for 2 h at −78° C. Then the reactionwas quenched with aqueous NaOH (2 M, 12 mL) and H₂O₂ (30%, 5 mL) andslowly warmed up to 23° C. over night. The resulting mixture wasextracted with Et₂O and the combined organic layer was washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. The crudematerial was passed through a short column to get the crude product. ¹HNMR (400 MHz, CDCl₃) δ 7.23 (2H, d, J=8.5 Hz), 6.86 (2H, d, J=8.5 Hz),5.81 (1H, m), 5.09 (2H, m), 4.79 (1H, d, J=6.8 Hz), 4.67 (1H, d, J=6.8Hz), 4.46 (1H, d, J=11.6 Hz), 4.43 (1H, d, J=11.6 Hz), 3.86 (2H, m),3.78 (3H, s), 3.62 (2H, m), 3.51 (1H, dd, J=6.6, 10.3 Hz), 3.44 (3H, s),3.36 (3H, s), 3.28 (1H, br s), 2.21 (2H, m), 1.72 (1H, m), 1.49 (1H, dt,J=14.5, 9, 3 Hz); ¹³C NMR (100 MHz, CDCl₃) δ 159.1, 134.7, 130.0, 129.2,117.4, 113.7, 96.8, 81.3, 77.2, 73.0, 70.3, 69.3, 58.4, 55.6, 55.2,42.1, 35.8;

Example 5

TBS ether (10) To a solution of the crude diastereomeric mixture (2.05g, 5.78 mmol) in DMF (15 mL) was added imidazole (720 mg, 10.6 mmol),DMAP (70 mg, 0.6 mmol) and TBSCl (1.06 g, 7.03 mmol) at 23° C. and thenthe mixture was stirred over night. To the resulting mixture was addedwater and EtOAc and the organic layer was washed with water and brine,dried over Na₂SO₄, concentrated in vacuo. Column chromatography providedthe pure diastereomeric isomer 10 (2.6 g, 74% over 4 steps, dr 94:6):[α]²³ _(D)-4.8 (c 2.69, CH₂Cl₂); IR (thin film, cm⁻¹) 3075, 2953, 2930,2895, 1856, 1612, 1513, 1249, 1099, 1039, 835; ¹H NMR (400 MHz, CDCl₃) δ7.24 (2H, d, J=8.6 Hz), 6.86 (2H, d, J=8.6 Hz), 5.82 (1H, m), 5.06 (1H,d, J=6.5 Hz), 5.03 (1H, s), 4.79 (1H, d, J=6.8 Hz), 4.65 (1H, d, J=6.8Hz), 4.45 (2H, s), 3.81 (2H, m), 3.80 (3H, s), 3.62 (1H, dd, J=4.9, 9.9Hz), 3.56 (1H, dd, J=6.2, 9.7 Hz), 3.47 (1H, m), 3.37 (3H, s), 3.36 (3H,s), 2.28 (1H, m), 2.23 (1H, m), 1.75 (1H, m), 1.64 (1H, m), 0.87 (9H,s), 0.05 (3H, s), 0.03 (3H, s); ¹³C NMR (100 MHz, CDCl₃) δ 159.0, 134.9,130.3, 129.2, 116.9, 113.6, 96.7, 77.5, 72.9, 69.8, 68.9, 58.0, 55.7,55.2, 41.9, 36.7, 25.8, 17.9, −4.4, −4.6; MS (ESI, m/z) [M+Na]⁺491.18;HRMS (ESI) [M+Na]⁺ calcd for C₂₅H₄₄O₆SiNa 491.2805, found 491.2806.OPMB

Example 6

Unsaturated ester (11) To the solution of thus obtained silyl ether 10(11 g, 23.5 mmol) in acetone/water (120 mL/15 mL) was added OsO₄ (2.5 w% in t-BuOH, 2.87 mL, 0.24 mmol) and NMO (3.31 g, 28.5 mmol) at 23° C.and the reaction mixture was stirred at that temperature for 5 h. Thesolid was removed and the filtrate was extracted with EtOAc. Thecombined organic layer was washed with water and brine, dried overNa₂SO₄ and concentrated in vacuo. To the solution of the crude diol inTHF/H₂O (100_(S-6) mL/25 mL) was added NaIO₄ (6.03 g, 28.2 mmol) and thereaction mixture was stirred at 23° C. for 3 h. The solid was removed byfiltration and the filtrate was extracted with EtOAc. The combinedorganic layer was washed with water and brine, dried over Na₂SO₄ andconcentrated in vacuo. The crude aldehyde was used without furtherpurification. To the solution of (O-cresol)₂P(O)CH₂CO₂Et (9.29 g, 29.4mmol) in THF (210 mL) was added NaI (3.45 g, 23 mmol) and NaH (60% inmineral oil, 1.03 g, 25.8 mmol) at 0° C. and it was stirred at 0° C. for10 min and cooled down to −78° C. To the resulting mixture was added thealdehyde in THF (50 mL) dropwise at −78° C. The reaction mixture wasstirred at −78° C. for 2 h and warmed up to −50° C. and then quenchedwith aqueous NH₄Cl and extracted with EtOAc. The combined organic layerwas washed with water and brine, dried over Na₂SO₄ and concentrated invacuo. Column chromatography provided the Z isomer 11 (9.9 g, 78%) and Eisomer (1.4 g, 11%). Z isomer: [α]²³ _(D) −9.5 (c 2.1, CH₂Cl₂); IR (thinfilm, cm⁻¹) 2953, 2930, 2897, 2857, 1718, 1514, 1250, 1180, 1098, 1039;¹H NMR (400 MHz, CDCl₃) δ 7.24 (2H, d, J=8.5 Hz), 6.86 (2H, d, J=8.5Hz), 6.37 91H, dt, J=11.6, 7.0 Hz), 5.84 (1H, d, J=m 11.6 Hz), 4.78 (1H,d, J=6.8 Hz), 4.64 (1H, d, J=6.8 Hz), 4.45 (2H, s), 4.14 (2H, q, J=7.2Hz), 3.96 (1H, m), 3.79 (3H, s), 3.61 (1H, dd, J=4.7, 9.8 Hz), 3.55 (1H,dd, J=6.3, 9.8 Hz), 3.46 (1H, m), 3.37 (3H, s), 3.36 (3H, s), 2.97 (1H,m), 2.82 (1H, m), 1.68 (2H, m), 1.27 (3H, t, J=7.3 hz), 0.86 (9H, s),0.05 (3H, s), 0.04 (3H, s); ¹³CNMR (100 MHz, CDCl₃) δ 166.2, 159.0,146.2, 130.3, 129.1, 121.0, 113.6, 96.6, 77.4, 76.5, 72.9, 69.7, 68.4,59.7, 58.2, 55.7, 55.2, 37.2, 36.1, 25.7, 17.9, 14.2, −4.5, −4.7; MS(ESI, m/z) [M+Na]⁺563.19; HRMS (ESI) [M+Na]⁺ calcd for C₂₈H₄₈O₈SiNa563.3016, found 563.3021.

Example 7

Acetonide (12) To the solution of the Z unsaturated olefin 11 (9.89 g,18.3 mmol) in t-BuOH/H₂O (92 mL/92 mL) was added AD-mix-α (25.7 g) andCH₃SO₂NH₂ (1.74 g) at 0° C. The reaction mixture was stirred at that 0°C. for 4 days and then quenched with NaHSO₃ and extracted with EtOAc.The combined organic layer was washed with water and brine, dried overNa₂SO₄ and concentrated in vacuo. The crude material was passed througha short silica gel column to provide the crude diastereomeric mixture(10.2 g, 97%), which could be separated in next step. To the solution ofcrude diol in CH₂Cl₂ (100 mL) was added PPTS (335 mg, 1.33 mmol) and2-methoxypropene (4.4 mL, 46 mmol) at 23° C. and the reaction mixturewas stirred for 30 min. The reaction solvent was removed in vacuo andpurification by column chromatography provided the major product 12(8.71 g) and minor product (1.17 g, overall 95%). The major isomer:[α]²³ _(D)+20.0 (c 1.89, CH₂Cl₂); IR (thin film, cm⁻¹) 2982, 2953, 2932,2896, 2856, 1757, 1513, 1463, 1249, 1099, 1039, 837; ¹H NMR (400 MHz,CDCl₃) δ 7.23 (2H, d, J=8.5 Hz), 6.86 (2H, d, J=8.6 Hz), 4.76 (1H, d,J=6.8 Hz), 4.64 (1H, d, J=6.8 Hz), 4.53 (1H, s), 4.50 (1H, m), 4.44 (2H,s), 4.24-4.14 (2H, m), 4.06 (1H, m), 3.79 (3H, s), 3.76 (1H, m), 3.60(1H, dd, J=4.5, 10.0 Hz), 3.50 (1H, dd, J=6.5, 9.9 Hz), 3.43 (1H, m),3.36 (3H, s), 3.35 (3H, s), 1.71-1.64 (3H, m), 1.58 (3H, s), 1.51 (1H,m), 1.35 (3H, s), 1.25 (3H, t, J=7.2 hz), 0.88 (9H, s), 0.07 (3H, s),0.05 (3H, s); ¹³C NMR (100 MHz, CDCl₃) δ 170.4, 159.0, 130.2, 129.1,113.6, 110.4, 96.5, 77.2, 76.9, 74.1, 72.9, 69.5, 66.4, 60.8, 58.5,55.6, 55.2, 39.0, 37.6, 27.0, 25.8, 25.7, 17.9, 14.1, −4.3, −4.9; MS(ESI, m/z) [M+Na]⁺637.21; HRMS (ESI) [M+Na]⁺ calcd for C₃₁H₅₄O₁₀SiNa637.3384, found 637.3392.

Example 8

Enone (2) To the solution of thus obtained ester 12 (1.47 g, 2.39 mmol)in CH₂Cl₂ (30 mL) was added DIBAL-H (1M in CH₂Cl₂, 2.5 mL, 2.5 mmol)dropwise at −78° C. The resulting mixture was stirred at thattemperature for 1 h and quenched with NH₄Cl. The solid was removed byfiltration and the filtrate was extracted with EtOAc. The combinedorganic layer was washed with brine, dried over Na₂SO₄, and concentratedin vacuo. The crude material was passed through a short silica gel padto provide the crude aldehyde, which was then dissolved in THF (50 mL).To the solution was added isopropenylmagnesium bromide (0.5 M in THF,26.9 mL, 13.4 mmol) dropwise at 0° C. and kept at 0° C. for 15 minbefore quenched with aqueous NH₄Cl. The resulting mixture was extractedwith EtOAc and the combined organic layer was washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. Columnchromatography provided the diastereomeric mixture (1.21 g, 86%). To thesolution of the obtained alcohol mixture (7.02 g, 11.4 mmol) in CH₂Cl₂(40 mL) was added Dess-Martin periodinane (5.83 g, 13.7 mmol) and NaHCO₃(3.46 g, 41.2 mmol) at 23° C. The reaction mixture was stirred for 30min and quenched with saturated aqueous Na₂S₂O₃ (20 mL) and saturatedaqueous NaHCO₃ (30 mL). The aqueous layer was extracted with Et₂O andthe combined organic layer was washed with water and brine, dried overNa₂SO₄ and concentrated in vacuo. Column chromatography provided theenone 2 (6.28 g, 90%). [α]²³ _(D)+29.3 (c 2.4, CH₂Cl₂); IR (thin film,cm⁻¹) 2953, 2931, 2894, 2856, 1693, 1513, 1378, 1249, 1100, 1073, 835;¹HNMR (400 MHz, CDCl₃) δ 7.22 (2H, d, J=8.5 Hz), 6.85 (2H, d, J=8.6 Hz),5.88 (1H, s), 5.84 (1H, d, J=1.1 Hz), 5.29 (1H, d, J=7.1 Hz), 4.74 (1H,d, J=6.8 Hz), 4.62 (1H, d, J=6.8 Hz), 4.58 (1H, m), 4.43 (2H, s), 4.01(1H, m), 3.78 (3H, s), 3.73 (1H, dt, J=6.7, 4.1 Hz), 3.58 (1H, dd,J=4.2, 9.9 Hz), 3.48 (1H, dd, J=6.6, 9.9 Hz), 3.37 (1H, m), 3.34 (3H,s), 3.32 (3H, s), 1.87 (3H, s), 1.60 (1H, m), 1.58 (3H, s), 1.40 (1H,m), 1.37 (3H, s), 0.87 (9H, s), −0.07 (3H, s), −0.04 (3H, s); ¹³C NMR(100 MHz, CDCl₃) δ 197.0, 159.0, 144.0, 130.2, 129.1, 125.3, 113.6,109.6, 96.6, 78.6, 77.2, 76.9, 74.5, 72.9, 69.5, 66.3, 58.5, 55.6, 55.1,39.0, 38.1, 27.3, 25.8, 25.5, 17.9, 17.8, −4.3, −4.9; MS (ESI, m/z)[M+Na]⁺633.16; HRMS (ESI) [M+Na]⁺ calcd for C₃₂H₅₄O₉SiNa 633.3435, found633.3433.

Example 9

TES ether (14) To the solution of the alcohol 5 (390 mg, 1.41 mmol) inCH₂Cl₂ (15 mL) was added 2,6-lutidine (0.63 mL, 5.36 mmol) and TESOTf(0.80 mL, 3.55 mmol). The resulting mixture was stirred for 5 min andwater and EtOAc was added. The organic layer was washed with water andbrine, dried over Na₂SO₄ and concentrated in vacuo. The crude materialwas passed through a silica gel pad to provide the crude silyl ether(495 mg, 90%). ¹H NMR (400 MHz, CDCl₃) δ 7.34 (4H, m), 7.28 (1H, m),5.81 (1H, m), 5.13-5.07 (2H, m), 4.92 (1H, d, J=10.2 Hz), 4.54 (1H, dd,J=5.0, 8.1 Hz), 4.51 (2H, s), 3.32 (3H, m), 2.57 (1H, m), 2.37 (1H, m),2.16 (1H, m), 1.71 (3H, d, J=1.0 Hz), 1.66 (1H, m), 1.23 (2H, m), 0.94(9H, t, J=7.8 Hz), 0.85 (3H, t, J=7.5 Hz), 0.56 (6H, q, J=7.8 Hz); ¹³CNMR (100 MHz, CDCl₃) δ 139.2, 138.6, 135.8, 128.2, 127.5, 127.4, 126.7,116.3, 73.5, 73.0, 70.6, 41.5, 39.1, 25.3, 17.9, 11.6, 6.8, 4.7.

Example 10

Alcohol (15) The TES ether was dissolved in t-BuOH/Acetone/H₂O (4 mL/4mL/1 mL). To the solution was added NMO (0.41 g, 3.53 mmol) and OsO₄(2.5 w % in t-BuOH, 0.88 mL, 0.074 mmol) at 0° C. and it was stirred atthis temperature for 2 h before being quenched with aqueous NaHSO₃. Theresulting mixture was extracted with EtOAc and the organic layer waswashed with brine, concentrated in vacuo and dissolved in CH₂Cl₂ (10mL). To the solution was added pyridine (0.3 mL, 3.7 mmol) and Pb(OAc)₄(0.66 g, 1.49 mmol) at 23° C. and stirred for 30 min. The solid wasremoved by filtration and the filtrate was extracted with EtOAc. Theorganic layer was washed with water and brine, dried over Na₂SO₄ andconcentrated in vacuo. The crude material was passed through a silicagel pad to give crude aldehyde (374 mg, 74%, 2 steps). To the solutionof (−)-Ipc₂BOMe (788 mg, 2.49 mmol) in Et₂O (12 mL) was addedallylmagnesium bromide (1 M in Et₂O, 2.22 mL, 2.22 mmol) dropwise at 0°C. The resulting mixture was warmed up to 23° C. over 2 h. The solid wasremoved by filtration and the filtrate was cooled down to −78° C. To thefiltrate was added thus obtained crude aldehyde in Et₂O (5 mL) viacannula at −78° C. over 5 min, and the mixture was stirred for 2 h at−78° C. and then quenched with buffer (pH=7) and H₂O₂ (30%, 5 mL). Theresulting mixture was slowly warmed up to 23° C. over night andextracted with Et₂O. The combined organic layer was washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. Columnchromatography provided major isomer (374 mg, 83%, dr 5:1).

Example 11

Methyl ether (16) To the solution of the thus made homoallylic alcohol(165 mg, 0.38 mmol) in CH₂Cl₂ (3 mL) was added proton sponge (333 mg,1.56 mmol) and Me₃OBF₄ (173 mg, 1.17 mmol) at 23° C. The reactionmixture was stirred for 3 h and the solid was removed by filtration. Thefilter cake was washed with hexane and the filtrate was washed withNaHCO₃, water and brine. The organic phase was dried over Na₂SO₄ andconcentrated in vacuo. Column chromatography provided methyl ether 13(153 mg, 93%): [α]²³ _(D) −35.3 (c 1.21, CH₂Cl₂); IR (thin film, cm⁻¹)3075, 3066, 3029, 2955, 2936, 2876, 2825, 1454, 1239, 1089, 1005, 741;¹H NMR (400 MHz, CDCl₃) δ7.34 (4H, m), 7.27 (1H, m), 5.86 (1H, m),5.13-5.07 (2H, m), 4.94 (1H, d, J=10.1 Hz), 4.60 (1H, dd, J=4.6, 8.8Hz), 4.50 (2H, s), 3.33 (3H, m), 3.32 (3H, s), 2.56 (1H, m), 2.38 (1H,m), 2.23 (1H, m), 1.94 (1H, ddd, J=4.9 8.9 13.8 Hz), 1.69 (3H, d, J=1.0Hz), 1.62 (1H, m), 1.44 (1H, ddd, J=4.5 7.5, 12.2 Hz), 1.24 (1H, m),0.92 (9H, t, J=8.0 Hz), 0.84 (3H, t, J=7.4 Hz), 0.56 (6H, q, J=7.9 Hz);¹³C NMR (100 MHz, CDCl₃) δ 138.9, 138.6, 134.7, 128.2, 127.5, 127.4,127.2, 116.9, 77.4, 73.3, 73.0, 67.5, 55.9, 40.1, 39.0, 37.4, 25.3,17.8, 11.4, 6.9, 4.8; MS (ESI, m/z) [M+Na]⁺469.21; HRMS (ESI) [M+Na]⁺calcd for C₂₇H₄₆O₃SiNa 469.3114, found 469.3123.

Example 12

Aldehyde (3) To the solution of the methyl ether 16 (133 mg, 0.30 mmol)in t-BuOH/Acetone/H₂O (1.0 mL/1.0 mL/0.25 mL) was added NMO (0.07 g,0.60 mmol) and OsO₄ (2.5 w % in t-BuOH, 0.15 mL, 0.013 mmol) at 0° C.and it was stirred at this temperature for 2 h before quenched withaqueous NaHSO₃. The resulting mixture was extracted with EtOAc and theorganic layer was washed with brine, concentrated in vacuo and dissolvedin CH₂Cl₂ (3 mL). To the solution was added pyridine (0.068 mL, 0.75mmol) and Pb(OAc)₄ (145 mg, 0.052 mmol) at 23° C. and stirred for 30min. The solid was removed by filtration and the filtrate was extractedwith EtOAc. The organic layer was washed with water and brine, driedover Na₂SO₄ and concentrated in vacuo. The crude material was purifiedby column chromatography to give the aldehyde 3 (100 mg, 75%, 2 steps):[α]²³ _(D)-40.1 (c 1.16, CH₂Cl₂); IR (thin film, cm⁻¹) 3029, 2956, 2935,2876, 1727, 1455, 1085, 1005, 742; ¹H NMR (400 MHz, CDCl₃) δ 9.80 (1H,t, J=1.9 Hz), 7.36-7.25 (5H, m), 4.94 (1H, d, J=10.3 Hz), 4.58 (1H, dd,J=3.6, 9.1 Hz), 4.49 (2H, s), 3.82 (1H, m), 3.34 (3H, s), 3.32 (2H, m),2.64 (1H, ddd, J=1.6, 4.1, 16.3 Hz), 2.57 (1H, ddd, J=2.7, 7.6, 16.5Hz), 2.51 (1H, m), 2.08 (1H, m), 1.71 (3H, s), 1.64 (1H, m), 1.45 (1H,ddd, J=3.6, 8.2, 13.7 Hz), 1.21 (1H, dt, J=13.6, 7.7 Hz), 0.92 (9H, t,J=8.0 Hz), 0.82 (3H, t, J=7.7 Hz), 0.56 (6H, q, J=7.7 Hz); ¹³C NMR (100MHz, CDCl₃) δ 201.4, 138.6, 138.5, 128.2, 127.5, 127.4, 73.7 73.3, 73.0,67.1, 56.2, 47.7, 40.1, 39.3, 25.3, 17.9, 11.6, 6.8, 4.7.

Example 13

Aldol product (17) To the solution of the enone 2 (645 mg, 1.06 mmol) inEt₂O (200 mL) was added L-Selectride (lithium tri-sec-butylborohydride,1.0 M in THF, 1.1 mL, 1.1 mmol) at −78° C. and the reaction was kept atthat temperature for 10-15 min. To the solution was added thus obtainedaldehyde 3 (520 mg, 1.2 mmol) in Et₂O (20 mL) at −78° C. The reactionmixture was stirred at −78° C. for 1 h and quenched with NH₄Cl. Theorganic layer was separated and the aqueous layer was extracted withEtOAc. The combined organic layer was washed with water and brine, driedover Na₂SO₄ and concentrated in vacuo. Column chromatography providedthe major isomer 17 and minor isomer (823 mg and 206 mg respectively,92%, dr 4:1). Major isomer: [α]²³ _(D)−13.9 (c 1.15, CH₂Cl₂); IR (thinfilm, cm⁻¹) 2956, 2936, 2879, 2859, 1714, 1514, 1463, 1379, 1249, 1098,1078, 1038, 836; ¹H NMR (400 MHz, CDCl₃) δ 7.30-7.25 (5H, m), 7.22 (2H,d, J=8.5 Hz), 6.84 (2H, d, J=8.5 Hz), 5.12 (1H, d, J=6.7 Hz), 4.92 (1H,d, J=10.1 Hz), 4.75 (1H, d, J=6.7 Hz), 4.63 (1H, d, J=6.8 Hz), 4.54 (2H,m), 4.46 (2H, s), 4.43 (2H, s), 4.01 (2H, m), 3.75 (3H, s), 3.59 (2H,dd, J=4.3, 10.0 Hz), 3.48 (1H, dd, J=6.7, 9.9 Hz), 3.39 (1H, m), 3.35(6H, s), 3.30 (2H, m), 3.29 (3H, s), 2.51 (1H, m), 2.05 (1H, m), 1.69(3H, s), 1.64-1.61 (4H, m), 1.57-1.42 (2H, m), 1.50 (3H, s), 1.33 (3H,s), 1.23 (2H, m), 1.18 (3H, s), 1.10 (3H, s), 0.94 (1H, m), 0.89 (9H, t,J=8.0 Hz), 0.87 (12H, s), 0.83 (3H, t, J=7.5 Hz), 0.54 (6H, q, J=8.0Hz), 0.08 (3H, s), 0.05 (3H, s); ¹³C NMR (100 MHz, CDCl₃) δ 210.2,159.0, 138.9, 138.5, 130.2, 129.1, 128.2, 127.4, 127.3, 127.2, 113.6,109.1, 96.6, 79.2, 77.3, 76.9, 76.5, 74.3, 73.2, 73.0, 72.9, 72.8, 69.5,67.3, 66.4, 58.5, 56.2, 55.6, 55.0, 51.3, 39.14, 39.05, 33.3, 27.6,25.8, 25.3, 20.8, 18.8, 17.94, 17.87, 11.6, 6.8, 4.7, −4.3, −4.9; MS(ESI, m/z) [M+Na]⁺1083.32; HRMS (ESI) [M+Na]⁺ calcd for C₅₈H₁₀₀O₁₃Si₂Na1083.6600, found 1083.6611.

Example 14

Alcohol (18) To the solution of the major aldol product 17 (30 mg, 0.028mmol) in THF/H₂O (9:1, 1.8 mL) was added DDQ (2 mg, 0.01 mmol). Thereaction mixture was stirred for 3 h at 23° C. and CH₂Cl₂ (7 mL), buffer(pH=7, 1.2 mL) and DDQ (30 mg, 0.13 mmol) was added. The resultingmixture was stirred for 8 h at 23° C. and aqueous NaHCO₃ was added. Theorganic layer was separated and the aqueous layer was extracted withEtOAc. The combined organic phase was washed with dilute aqueous NaHCO₃,water and brine, dried over Na₂SO₄ and concentrated in vacuo.Column_(S-11) chromatography gave the alcohol 18 (16 mg, 70%):: [α]²³_(D)+17.9 (c 1.3, CHCl₃); IR (thin film, cm⁻¹) 3443, 2956, 2928, 2855,1713, 1462, 1378, 1252, 1097, 1074, 836; ¹HNMR (400 MHz, CDCl₃) δ7.34-7.24 (5H, m), 5.10 (1H, d, J=6.6 Hz), 4.95 (1H, d, J=9.5 Hz), 4.71(1H, d, J=6.9 Hz), 4.65 (1H, d, J=6.9 Hz), 4.59 (1H, dd, J=4.8, 8.5 Hz),4.52 (1H, m), 4.47 (2H, m), 4.02-3.95 (2H, m), 3.69 (1H, m), 3.63-3.53(3H, m), 3.42-3.36 (2H, m), 3.40 (3H, s), 3.35 (3H, s), 3.32 (3H, s),3.12 (1H, t, J=9.1 Hz), 2.65 (1H, m), 2.02 (1H, m), 1.72 (3H, s),1.69-1.55 (6H, m), 1.50 (3H, s), 1.20 (2H, m), 1.18 (3H, s), 1.13 (1H,m), 1.09 (3H, s), 0.87 (9H, s), 0.82 (3H, t, J=7.4 Hz), 0.07 (3H, s),0.05 (3H, s); ¹³C NMR (100 MHz, CDCl₃) δ 210.6, 139.3, 137.6, 130.9,128.4, 127.7, 109.2, 97.3, 81.1, 79.1, 78.1, 77.9, 74.3, 73.7, 73.4,73.2, 66.3, 65.7, 62.4, 58.2, 56.9, 55.7, 51.4, 39.3, 38.9, 38.6, 36.5,33.5, 29.6, 27.6, 25.8, 24.7, 20.8, 18.9, 18.1, 17.9, 11.8, −4.3, −4.9;MS (ESI, m/z) [M+Na]⁺849.2; HRMS (ESI) [M+Na]⁺ calcd for C₄₄H₇₈O₁₂SiNa849.5160, found 849.5154.

Example 15

Macrolactone (19) To the solution of the alcohol 18 (17.3 mg, 0.021mmol) in CH₂Cl₂ (1.9 mL) was added 4 Å molecular sieve, NMO (2.2 mg,0.021 mmol) and TPAP (1.2 mg, 0.004 mmol) at 0° C. The resulting mixturewas stirred at 0° C. for 1 h. The solid was removed by filtration andEtOAc was added to the filtrate. The organic phase was washed withaqueous Na₂S₂O₃, water and brine, concentrated in vacuo and dissolved int-BuOH (3.3 mL). To the solution was added 2-methyl-2-butene (0.4 mL)and a solution of NaClO₂ (32 mg, 0.35 mmol) and NaH₂PO₄ (35 mg, 0.29mmol) in H₂O (3.3 mL). The resulting mixture was stirred at 23° C. for25 min. The organic layer was separated and the aqueous layer wasextracted with EtOAc. The combined organic phase was washed with waterand brine, dried over Na₂SO₄ and concentrated in vacuo. The crudeproduct was passed through a short silica gel column to obtain the crudeseco-acid 9 mg. To the solution of thus obtained seco-acid in toluene(2.4 mL) was added DIPEA (0.05 mL, 0.29 mmol) and 2,4,6-trichlorobenzoylchloride (18.8 ┌L, 0.12 mmol) at 23° C. The reaction was stirred for 15h at that temperature and was added dropwise to a solution of DMAP (22.4mg, 0.18 mmol) in toluene (25 mL) at 23° C. over 10 h. The resultingmixture was stirred at 23° C. for 36 h and water was added. The organiclayer was separated and the aqueous was extracted with EtOAc. Thecombined organic phase was wash with 0.18% HCl, water and brine, driedover Na₂SO₄ and concentrated in vacuo. Column chromatography providedproduct 19 (5.7 mg, 64%): [α]²³ _(D)−45.7 (c 0.88, CHCl₃); IR (thinfilm, cm⁻¹) 2956, 2929, 2856, 1730, 1463, 1379, 1256, 1095, 1076, 1027,971, 837; ¹H NMR (400 MHz, CDCl₃) δ 7.35-7.22 (5H, m), 5.78 (1H, d,J=8.0 Hz), 5.06 (1H, d, J=10.2 Hz), 4.90 (1H, d, J=6.9 Hz), 4.70 (1H,_(S-12)d, J=6.7 Hz), 4.68 (1H, d, J=6.7 Hz), 4.63 (1H, m), 4.53 (1H, d,J=12.1 Hz), 4.47 (1H, d, J=12.1 Hz), 4.08 (1H, m), 3.98 (1H, d, J=5.1Hz), 3.81 (1H, m), 3.59 (1H, dd, J=4.1, 9.3 Hz), 3.49 (1H, m), 3.40 (3H,s), 3.37 (3H, s), 3.34 (6H, s), 3.20 (1H, m), 2.75 (1H, m), 2.05 (1H,ddd, J=15.2, 5.5, 1.1 Hz), 1.97 (1H, ddd, J=15.2, 9.1, 1.3 Hz), 1.88(1H, ddd, J=14.5, 6.3, 4.3 Hz), 1.72 (2H, m), 1.66 (3H, s), 1.63 (1H,m), 1.59 (3H, s), 1.53 (1H, m), 1.42 (3H, s), 1.37 (3H, s), 1.32 (1H,m), 1.22 (1H, m), 1.18 (3H, s), 0.88 (9H, s), 0.84 (3H, t, J=7.5 Hz),0.09 (3H, s), 0.08 (3H, s); ¹³C NMR (100 MHz, CDCl₃) δ 216.1, 168.8,138.9, 134.0, 129.8, 128.1, 127.5, 127.2, 110.2, 96.4, 79.4, 77.4, 76.7,75.4, 74.0, 72.7, 70.6, 66.0, 58.4, 57.4, 56.2, 50.4, 39.5, 39.3, 38.5,37.9, 35.7, 29.6, 26.7, 25.8, 25.5, 25.2, 24.9, 21.0, 17.9, 11.6, −4.2,−5.0; MS (ESI, m/z)[M+Na]⁺845.24; HRMS (ESI) [M+Na]⁺ calcd forC₄₄H₇₄O₁₂SiNa 845.4847, found 845.4840.

Example 16

Methyl ether (20). The macrolactone 19 (13 mg, 0.016 mmol) was dissolvedin a mixture of THF (3.6 mL) and 1N HCl (3.6 mL). The resulting mixturewas stirred at 23° C. for 9 h. The aqueous layer was extracted withEtOAc and the combined organic phase was washed with aqueous NaHCO₃,water and brine, dried over Na₂SO₄ and concentrated in vacuo. The crudematerial was passed through a silica gel pad to give the crude productwhich was then dissolved in CH₂Cl₂. To the solution was added2,6-di-tert-butylpyridine (60 μL), Me₃OBF₄ (24 mg) at 0° C. The reactionwas stirred for 4 h and quenched with aqueous NaHCO₃. The organic phasewas separated and the aqueous layer was extracted with EtOAc. Combinedorganic phase was washed with water and brine, dried over Na₂SO₄ andconcentrated in vacuo. A simple silica gel column gave 8 mg crudeproduct, which may contains both the macrolactone form and thesemi-ketal form. MS (ESI, m/z) [M+Na]⁺705.29.

Example 17

Peloruside A. To the solution of the methyl ether (4 mg) in methanol (2mL) was added formic acid (0.1 mL) and a catalytic amount of 10% Pd/C at23° C. and the resulting mixture was stirred for 1 h. Celite® was addedand the solid was removed by filtration. The filtrate S-13 wasconcentrated in vacuo and was dissolved in THF/4N HCl (1.5 mL/1.5 mL)and was stirred for 3.5 h at 23° C. The reaction mixture was extractedwith EtOAc and the combined organic layer was washed with NaHCO₃, waterand brine, dried over Na₂SO₄ and concentrated in vacuo. Columnchromatography provided the (+)-Peloruside A (1) (1.6 mg, 50%, 2 steps)[α]²³ _(D)+15.1 (c 0.1, CH₂Cl₂); IR (thin film, cm⁻¹) 2957, 2923, 2852,1742, 1463, 1378, 1151, 1086, 1037, 722; ¹H NMR (500 MHz, CDCl₃)

36.79 (1H, br s), 5.69 (1H, d, J=10.6 Hz), 5.05 (1H, d, J=10.5 Hz), 4.91(1H, m), 4.54 (1H, br d, J=8.2 Hz), 4.47 (1H, s), 4.28 (1H, ddd, J=11.3,4.4, 2.5 Hz), 4.23 (1H, dd, J=10.6, 5.4 Hz), 4.02 (1H, d, J=2.8 Hz),3.99 (1H, m), 3.82 (1H, ddd, J=11.5, 5.0, 3.0 Hz), 3.65 (1H, br d,J=10.5), 3.48 (3H, s), 3.39 (3H, s), 3.36 (1H, m), 3.31 (3H, s), 3.01(1H, br s), 2.70 (1H, d, J=9.3 Hz), 2.62 (1H, m), 2.27 (1H, br s), 2.14(1H, m), 2.05 (1H, m), 1.79 (1H, ddd, J=12.5, 4.9, 2.5 Hz), 1.78 (1H,m), 1.68 (3H, d, J=1.1 Hz), 1.53 (1H, q, J=12.0 Hz), 1.46-1.40 (2H, m),1.17 (1H, m), 1.13 (3H, s), 1.10 (3H, s), 0.87 (3H, t, J=7.5 Hz); ¹³CNMR (125 MHz, CDCl₃) δ 174.0, 136.1, 131.2, 102.0, 78.3, 78.0, 76.0,73.9, 70.9, 70.3, 67.0, 66.9, 63.5, 59.1, 56.1, 55.7, 43.6, 43.4, 35.8,33.9, 32.6, 31.7, 24.7, 20.9, 17.5, 15.8, 12.3; MS (ESI, m/z)[M+Na]+571.17; HRMS (ESI) [M+Na]⁺ calcd for C₂₇H₄₈O₁₁Na 571.3094, found571.3102. IC₅₀ (P388 murine leukemia cells) 10 nM.

Example 18

epi-C-11 Peloruside A. epi-C-11 peloruside was prepared in an analogousmanner. ¹H NMR (500 MHz), δ 7.33 (br s, 1H), 5.60 (d, J=10.2 Hz, 1H),5.09 (d, J=10.4 Hz), 4.92 (br s, 1H), 4.31 (s, 1H), 4.29 (m, 1H), 4.14(m, 1H), 3.95 (d, J=2.7 Hz, 1H), 3.76 (ddd, J=2.9, 4.8, 9.4 Hz, 1H),3.67 (m, 2H), 3.54 (d, J=10.7 Hz, 1H), 3.47 (s, 3H), 3.41 (s, 3H), 3.37(s, 3H), 3.39-3.37 (m, 1H), 2.57 (m, 1H), 2.32 (dt, J=15.5, 11.1 Hz),2.08 (m, 2H), 2.01-1.96 (m, 2H), 1.80 (m, 2H), 1.76 (m, 3H), 1.57 (t,J=11.8 Hz, 2H), 1.43 (m, 1H), 1.40 (s, 3H), 1.33-1.25 (m, 1H), 1.19 (s,3H), 0.90 (t, J=7.3 Hz, 3H); ¹³C NMR (125 MHz), δ 173.4, 136.1, 131.0,102.5, 87.8, 83.1, 77.3, 75.1, 72.2, 70.7, 68.8, 67.0, 64.6, 57.1, 56.6,55.7, 43.5, 43.3, 39.3, 34.4, 33.3, 31.4, 29.7, 24.5, 23.5, 22.3, 17.5,12.3; IR (cm⁻¹) 3412, 2956, 2925, 2876, 2854, 1742, 1424, 1223, 1083,1000. epi-C-11 was equipotent to peloruside A (10 nM) in the cellproliferation assay.

Example 19

2-des-Hydroxy-Peloruside A. The 2-des-hydroxy compound was preparedusing procedures similar to those described herein. It gave an IC₅₀ of120 nM in the cell proliferation assay.

Example 20

2-des-Hydroxy-7-des-methoxy-7-hydroxy-Peloruside A. The2-des-hydroxy-7-hydroxy compound was prepared using procedures similarto those described herein. It gave an IC₅₀ of 320 nM in the cellproliferation assay.

Example 21

Open Chain Isomer Of 2-Des-Hydroxy-Peloruside A. The des-lactol isomerof 2-des-hydroxy-peloruside A was prepared using procedures similar tothose described herein. It gave an IC₅₀ of >5 μM in the cellproliferation assay.

What is claimed is:
 1. A process for forming a quaternary carbon betweena hydroxymethylene or alkoxymethylene carbon and a ketone (or hemiketalor ketal thereof) carbon or a hydroxymethylene reduction product of saidketone, the process comprising the steps of reacting a first compoundcontaining an α-alkyl-α,β-unsaturated ketone moiety with a source ofnucleophilic hydride, wherein the source of nucleophilic hydride islithium (sec-butyl)₃borohydride; and adding a second compound containingan aldehyde to form the quaternary carbon.
 2. A process of forming acompound wherein the compound contains a quaternary carbon between ahydroxymethylene or alkoxymethylene carbon and a ketone (or hemiketal orketal thereof) carbon or a hydroxymethylene reduction product of saidketone comprising the steps of reacting a first compound containing anα-alkyl-α,β-unsaturated ketone moiety with a source of nucleophilichydride, wherein the source of nucleophilic hydride is lithium(sec-butyl)₃borohydride; and adding a second compound containing analdehyde to form the quaternary carbon.
 3. The process of claim 2wherein the compound containing a quaternary carbon is a bryostatin, apeloruside, a mycalamide or an epothilone.
 4. The process of claim 2wherein the compound containing a quaternary is a macrolactone which hasthe formula

wherein: R¹ is

where * shows the point of attachment and R⁸ and R¹² are independentlyhydrogen, alkyl, heteroalkyl, arylalkyl, heteroarylalkyl, or an oxygenprotecting group; R², R³, R⁴, R⁵, R⁶ and R⁷ are each independentlyselected from the group consisting of hydrogen, alkyl, hydroxy, andalkoxy; and X¹ is hydrogen, hydroxy, alkoxy or together with the carbonto which it is attached forms a carbonyl or an oxime.
 5. The process ofclaim 4 wherein the macrolactone has the formula

wherein: R¹ is

where * shows the point of attachment; R², R³, R⁴, R⁵, R⁶ and R⁷ areeach independently selected from the group consisting of hydrogen,alkyl, hydroxy, and alkoxy; X¹ is hydrogen, hydroxy, alkyl, alkoxy ortogether with the carbon to which it is attached forms a carbonyl or anoxime; and R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl,arylalkyl, heteroarylalkyl, or an oxygen protecting group.
 6. Theprocess of claim 5 wherein the macrolactone has the formula

wherein R¹ is

where * shows the point of attachment; R², R³, R⁴, R⁵, R⁶ and R⁷ areeach independently selected from the group consisting of hydrogen,alkyl, hydroxy, and alkoxy; X¹ is hydrogen, hydroxy, alkyl, alkoxy ortogether with the carbon to which it is attached forms a carbonyl or anoxime; and R⁸ and R¹² are independently hydrogen, alkyl, heteroalkyl,arylalkyl, heteroarylalkyl, or an oxygen protecting group.
 7. Theprocess of claim 6 wherein R³ is methoxy and R⁷ is methoxy.
 8. Theprocess of claim 6 wherein R⁶ is hydrogen.
 9. The process of claim 6wherein R² is hydrogen.
 10. The process of claim 6 wherein R² ishydroxy; R⁴ is methoxy; R⁵ is hydroxy; R⁶ is hydroxy; X¹ is hydroxy; andR¹ is

where * shows the point of attachment and R⁸ and R¹² are independentlyhydrogen, alkyl, heteroalkyl, arylalkyl, heteroarylalkyl, or an oxygenprotecting group.
 11. The process of claim 6 wherein R² is hydroxy; R⁴is methoxy; R⁵ is hydroxy; R⁶ is hydrogen, hydroxy, alkyl, or alkoxy; X¹is hydroxy; and R¹ is 4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl.
 12. Theprocess of claim 6 wherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydroxy;R⁶ is hydroxy; X¹ is hydrogen, or epi-hydroxy, alkoxy, or alkyl, or X¹and the attached carbon form a carbonyl; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl.
 13. The process of claim 6wherein R² is hydrogen, hydroxy, or alkyl; R⁴ is methoxy; R⁵ is hydroxy;R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl.
 14. The process of claim 6wherein R² is hydroxy; R⁴ is hydrogen, hydroxy, alkoxy, or alkyl; R⁵ ishydroxy; R⁶ is hydroxy; X′ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl.
 15. The process of claim 6wherein R² is hydroxy; R⁴ is methoxy; R⁵ is hydrogen, hydroxy, alkoxy,or alkyl; R⁶ is hydroxy; X¹ is hydroxy; and R¹ is4-(R)-hydroxymethyl-hex-2-(Z)-ene-2-yl.
 16. The process of claim 1wherein alkyl is C₁-C₄ alkyl.
 17. The process of claim 16 wherein alkylis methyl.
 18. The process of claim 1 wherein a moiety of the followingformula

comprises the quaternary carbon formed.
 19. The process of claim 1wherein a moiety of the following formula

comprises the quaternary carbon formed.
 20. The process of claim 2wherein alkyl is C₁-C₄ alkyl.
 21. The process of claim 20 wherein alkylis methyl.
 22. The process of claim 2 wherein a moiety of the followingformula

comprises the quaternary carbon formed.
 23. The process of claim 2wherein a moiety of the following formula

comprises the quaternary carbon formed.